Skeletal regenerative medicine promises to mitigate age-related bone loss and support fracture healing but requires manipulation of biological properties of osteogenic progenitor cells. Osteogenic cells must sustain their phenotype during lineage-expansion yet undergo a number of mitotic cell cycles to generate the requisite number of cells for proper tissue-organization. The proposed study will examine fundamental mechanisms that ensure cells retain a post-mitotic molecular memory of the bone phenotype. The central hypothesis of our study is that key mRNAs for osteogenic factors are passed on to daughter cells during mitosis as a component of a non-genomic epigenetic mechanism and that selected microRNAs (miRs) attenuate the translation of these transmitted mRNAs. Our hypothesis is based on a robust set of preliminary data showing that the osteogenic master regulator Runx2 is controlled by mitotic miRs in osteoblastic cell lines. Based on these and other preliminary data, we will (i) characterize mitotic mRNAs and cognate miRs that suppress expression of lineage-specific factors in osteogenic cells, (ii) examine miR dependent changes in protein expression and lineage-plasticity in proliferating osteoblasts, and (iii) characterize the physiological role of mitosis-related miRs during skeletal development in vivo. Validation of this concept would (i) establish a major new dimension in cell cycle regulation, (ii) reveal a previously unrecognized function for miRs during mitosis, (iii) define a novel molecular mechanism for biological control of gene expression in lineage-committed cells, and (iv) identify specific miRs that are transmitted to osteogenic progeny cells upon mitosis ('mito-miRs'). From a molecular therapeutic perspective, these miRs permit generation of epigenomic agents that control cellular inheritance by targeting cell fate determining factors which mediate mesenchymal stem cell expansion and differentiation.